HiSense Mycoplasma PCR Detection Kit

HiSenseTM Mycoplasma PCR Detection Kit utilizes the PCR, which is established as the method of choice for highest sensitivity in the detection of mycoplasma, ureaplasma and acholeplasma contamination in cell culture and other cell culture derived biologicals. The primers are specific to the highly conserved 16S ribosomal RNA sequences in the mycoplasma genome. This allows for detection of all mycoplasma species including Acholeplasma laidlawii, M. arginini, M. fermentans, M. gallisepticum, M. genitalium, M. hominis, M hyorhinis, M. orale, M. pneumoniae, Spiroplasma citri and U. urealyticum. All mycoplasma species including 8 genus and about 209 species can be detected simultaneously. The “European Pharmacopoeia” and Guideline of “Korea Ministry of Food and Drug Safety” (KMFDS) recommend checking for unspecific detection of Clostridium, Lactobacilllus, and Streptococcus. The 16S ribosomal RNA of other bacteria such as E.coli, Clostridium, Lactobacillus, Sterptococcus, and plant and animal cells is not amplified.

Cat. No HDEP-25, HDEP-50, HDEP-100 (Number of Sample)
Storage -20°C
Intended Use detecting mycoplasma contamination in pharmaceutical manufacturing facilities and biopharmaceuticals
Method Conventional PCR
Sample Material Template DNA prepared by boiling of culture supernatant or by genomic DNA extraction
Sensitivity 1~10 fg/reaction
HiSenseEP PCR Detection Kit Mannual(Ver4.0) 2022-07-13 18:30:32 765,45 KB

Characteristics of CellSafe’s Mycoplasma Detection Kit

CellSafe’s Mycoplasma Detection kits utilize the PCR technique which is established as the method of choice for the highest sensitivity in the detection of mycoplasma contamination in cell culture and other cell culture derived biologicals. Detection of M.orale, M.hyorhinis, M.arginini, M.fermentans, M.hominis and Acholeplasma laidlawii usually encountered as contaminants in cell cultures. Furthermore, these kits can detect M.pneumoniae, M.salivarium, M.genitalium, M.penentrans and Ureaplasma species. The primers are specific to the highly conserved 16s rRNA coding region and the kits do not detect other bacteria and cell-lines. All kits include Internal Amplification Control (IAC) providing validity of test results. They are user firendly, ready-to-use, optimized PCR master mix type kits.

 

Kit Specificity

The HiSenseTM Mycoplasma PCR Detection Kit detects Mycoplasma species simply, reliably, and rapidly. To detect the presence of these microorganisms, the assay uses the polymerase chain reaction (PCR) to amplify a target unique to a wide variety of mycoplasmas. The kit can detect more than 209 different Mycoplasma species, including Acholeplasma laidlawii and Spiroplasma citri. The kit does not detect other genera or cell-line DNA.

 

 

                                   


Kit Sensitivity

The sensitivity of the PCR using this kit is 1 to 10 copies of the target DNA per reaction. Sensitivity of the assay in real culture samples depends on the quality of the sample preparation.

 

 

Preventing Cross-Contamination

Take precautions to minimize the potential for carryover of nucleic acids from one experiment to the next. Use separate work areas and pipettors for template preparation and PCR setup steps. Use the PCR grade water delivered with the kit, aerosol-preventive filter tips and gloves.
The 2X PCR master mix contains dUTP instead of dTTP. When dUTP replaces dTTP in PCR amplification, UNG treatment (Uracil-N-glycosylase, not provided in this kit) can prevent the subsequent reamplification of dU-containing PCR products. UNG acts on single- and double-stranded dU-containing DNA by hydrolysis of uracil-glycosidic bonds at dU-containing DNA sites. When this strategy is put to use, carryover contamination will be eliminated while template DNA (DNA containing T) will be left intact.